• 생명과학회지
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• 제28권11호
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• pp.1332-1338
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• 2018

뇌신경 질환의 주요 원인이 되는 것으로 알려진 미세아교세포의 과도한 활성화에 의한 신경염증반응에서 풀무치 에탄올 추출물이 미세아교세포의 염증 반응에 미치는 영향을 검토하였다. 미세아교세포의 활성화를 유도하기 위해 LPS를 사용하였으며, LPS 처리에 의해 신경염증반응의 지표인 NO의 생성량과 이들을 조절하는 iNOS, COX-2의 발현이 증가됨을 확인 할 수 있었다. 그러나 풀무치 에탄올 추출물을 1시간 전처리 한 후 LPS를 처리한 경우 추출물의 농도에 의존적으로 이들의 발현량이 현저히 감소되는 것을 확인 하였다. 또한 LPS 처리로 인해 분비되는 염증성 cytokine들의 생성량도 풀무치 에탄올 추출물에 의해 현저히 억제 됨을 확인 할 수 있었다. 따라서 본 연구의 결과는 미세아교세포의 과도한 활성화로 인해 발생되는 뇌 신경질환의 치료 소재로서 풀무치 에탄올 추출물의 활성 가능성을 제시하였다.

• BMB Reports
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• 제54권11호
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• pp.557-562
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• 2021

Microglial activation is closely associated with neuroinflammatory pathologies. The nucleotide-binding and oligomerization domain-like receptor containing a pyrin domain 3 (NLRP3) inflammasomes are highly organized intracellular sensors of neuronal alarm signaling. NLRP3 inflammasomes activate nuclear factor kappa-B (NF-κB) and reactive oxygen species (ROS), which induce inflammatory responses. Moreover, NLRP3 dysfunction is a common feature of chronic inflammatory diseases. The present study investigated the effect of a novel thiazol derivative, N-cyclooctyl-5-methylthiazol-2-amine hydrobromide (KHG26700), on inflammatory responses in lipopolysaccharide (LPS)-treated BV-2 microglial cells. KHG26700 significantly attenuated the expression of several pro-inflammatory cytokines, including tumor necrosis factor-α, interleukin-1β, and interleukin-6, in these cells, as well as the LPS-induced increases in NLRP3, NF-κB, and phospho-IkBα levels. KHG26700 also suppressed the LPS-induced increases in protein levels of autophagy protein 5 (ATG5), microtubule-associated protein 1 light chain 3 (LC3), and beclin-1, as well as downregulating the LPS-enhanced levels of ROS, lipid peroxidation, and nitric oxide. These results suggest that the anti-inflammatory effects of KHG26700 may be due, at least in part, to the regulation of the NLRP3-mediated signaling pathway during microglial activation.

• The Korean Journal of Physiology and Pharmacology
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• 제19권3호
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• pp.219-228
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• 2015

Excessive microglial activation and subsequent neuroinflammation lead to synaptic loss and dysfunction as well as neuronal cell death, which are involved in the pathogenesis and progression of several neurodegenerative diseases. Thus, the regulation of microglial activation has been evaluated as effective therapeutic strategies. Although dieckol (DEK), one of the phlorotannins isolated from marine brown alga Ecklonia cava, has been previously reported to inhibit microglial activation, the molecular mechanism is still unclear. Therefore, we investigated here molecular mechanism of DEK via extracellular signal-regulated kinase (ERK), Akt and nicotinamide adenine dinuclelotide phosphate (NADPH) oxidase-mediated pathways. In addition, the neuroprotective mechanism of DEK was investigated in microglia-mediated neurotoxicity models such as neuron-microglia co-culture and microglial conditioned media system. Our results demonstrated that treatment of anti-oxidant DEK potently suppressed phosphorylation of ERK in lipopolysaccharide (LPS, $1{\mu}g/ml$)-stimulated BV-2 microglia. In addition, DEK markedly attenuated Akt phosphorylation and increased expression of $gp91^{phox}$, which is the catalytic component of NADPH oxidase complex responsible for microglial reactive oxygen species (ROS) generation. Finally, DEK significantly attenuated neuronal cell death that is induced by treatment of microglial conditioned media containing neurotoxic secretary molecules. These neuroprotective effects of DEK were also confirmed in a neuron-microglia co-culture system using enhanced green fluorescent protein (EGFP)-transfected B35 neuroblastoma cell line. Taken together, these results suggest that DEK suppresses excessive microglial activation and microglia-mediated neuronal cell death via downregulation of ERK, Akt and NADPH oxidase-mediated pathways.

• 대한본초학회지
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• 제28권6호
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• pp.79-86
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• 2013

Objectives : The purpose of this study is to investigate neuroprotective effects and molecular mechanisms of luteolin against ${\beta}$-amyloid ($A{\beta}_{25-35}$)-induced oxidative cell death in BV-2 cells. Methods : The protective effects of luteolin against $A{\beta}_{25-35}$-induced cytotoxicity and apoptotic cell death were determined by MTT dye reduction assay and TUNEL staining, respectively. The apoptotic cell death was further analyzed by measuring mitochondrial transmembrane potential and expression of pro- and/or anti-apoptotic proteins. To elucidate the molecular mechanisms underlying the protective effects of luteolin, intracellular accumulation of reactive oxygen species, oxidative damages, and expression of antioxidant enzymes were examined. Results : Luteolin pretreatment effectively attenuated $A{\beta}_{25-35}$-induced apoptotic cell death indices such as DNA fragmentation, dissipation of mitochondrial transmembrane potential, increased Bax/Bcl-2 ratio, and activation of c-Jun N-terminal kinase and caspase-3 in BV-2 cells. Furthermore, $A{\beta}_{25-35}$-induced intracellular formation of reactive oxygen species and subsequent oxidative damages such as lipid peroxidation and depletion of endogenous antioxidant glutathione were suppressed by luteolin treatment. The neuroprotective effects of luteolin might be mediated by up-regulation of cellular antioxidant defense system via up-regulation of ${\gamma}$-glutamylcysteine ligase, a rate-limiting enzyme in the glutathione biosynthesis and superoxide dismutase, an enzyme involved in dismutation of superoxide anion into oxygen and hydrogen peroxide. Conclusions : These findings suggest that luteolin has a potential to protect against $A{\beta}_{25-35}$-induced neuronal cell death and damages thereby exhibiting therapeutic utilization for the prevention and/or treatment of Alzheimer's disease.

• 동의생리병리학회지
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• 제20권5호
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• pp.1200-1210
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• 2006

This experiment was designed to investigate the effect of the CMT and MCMT hot water extract & ultra-fine powder on microglia and memory deficit model. The effects of the CMT and MCMT hot water extract on expression of IL-l${\beta}$, IL-6, TNF-${\alpha}$, NOS-II, COX-2, IL-10, TGF-${\beta}$1 mRNA and production of IL-lP, IL-6, TNF-a, NO, ROS in BV2 microglial cell line treated by lipopolysacchaide(LPS) ; serum glucose, uric acid, AChE activity of the memory deficit mice induced by scopolamine , behavior of the memory deficit mice induced by scopolamine and were investigated, respectively. The CMT and MCMT hot water extract suppressed the expression of IL-1${\beta}$, IL-6, TNF-${\alpha}$, NOS-II, COX-2 mRNA, production of IL-l${\beta}$, IL-6, TNF-${\alpha}$, NO, ROS and increased the expression of IL-10, TGF-${\beta}$l mRNA in BV2 microglial cell line treated by LPS. The MCMT hot water extract & ultra-fine powder increased glucose, decreased uric acid and AChE significantly in the serum of the memory deficit mice induced by scopolamine. The CMT and MCMT hotwater extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. According to the above result, it is suggested that the CMT and MCMT hot water extract & ultra-fine powder might be usefully applied for prevention and treatment of dementia.

• 동의생리병리학회지
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• 제20권4호
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• pp.997-1008
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• 2006

This experiment was designed to investigate the effect of the CMT and PCMT hot water extract & ultra-fine powder on microglia and memory deficit model. The effects of the CMT and PCMT hot water extract on expression of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NOS-II, COX-2, IL-10, $TGF-{\beta}1$ mRNA and production of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NO, ROS in BV2 microglial cell line treated by lipopolysacchaide(LPS) , serum glucose, uric acid, AChE activity of the memory deficit mice induced by scopolamine , behavior of the memory deficit mice induced by scopolamine and were investigated, respectively. The CMT and PCMT hot water extract suppressed the expression of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NOS-11, COX-2 mRNA, production of $IL-l{\beta},\;IL-6,\;TNF-{\alpha}$, NO, ROS and increased the expression of IL-10, $TGF-{\beta}1$ mRNA in BV2 microglial cell line treated by LPS. The PCMT hot water extract & ultra-fine powder increased glucose, decreased uric acid and AChE significantly in the serum of the memory deficit mice induced by scopolamine. The CMT and PCMT hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. According to the above result, it is suggested that the CMT and PCMT hot water extract & ultra-fine powder might be usefully applied for prevention and treatment of dementia.

• 동의생리병리학회지
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• 제24권3호
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• pp.463-469
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• 2010

This research is performed to obtain positive evidences of Mori cortex, a kind of oriental medicinal herbs, in the cellular levels. The extracts of M. cortex have shown anti-inflammatory effects against cutaneous inflammation and clinical effects on pulmonary asthma and congestion in oriental medicine. Thus BV2 cells were chosen because microglia are considered as the main immunocompetent cells in the central nervous system. Lipopolysaccharide (LPS)-induced microglial activation of cultured BV2 cells and subsequent release of nitric oxide (NO) and Prostaglandin E2 (PGE2) were effectively suppressed by methylene chloride extract of Morus alba L. (MEMA). From the inflammation-mediated mRNA and protein analyses, we showed that inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-$1{\beta}$ (IL-$1{\beta}$) and tumor necrosis alpha (TNF-${\alpha}$) induced by LPS were markedly decreased by MEMA treatment. From the observation of nuclear factor-kB (NF-${\kappa}B$) which is controlling and mediating inflammation through COX-2 and iNOS, there showed that p65, a subunit of NF-${\kappa}B$, was increased in nuclear and $I{\kappa}B$, a competitor of NF-${\kappa}B$, was recovered in cytosol after MEMA treatment. These are corresponding with results of iNOS, COX-2, IL-$1{\kappa}$ and TNF-${\alpha}$, and confirm some suppressive effect against transcriptional activation of NF-${\kappa}B$. In conclusion, the anti-inflammatory action of M. cortex against BV2 microglia cells is expected to protect nerve tissues through suppression of neuronal inflammation in various neurodegenerative diseases.

• 동의신경정신과학회지
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• 제19권3호
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• pp.55-68
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• 2008

Background: Microglia produces a barrage of factors (IL-l, TNF-$\alpha$, NO, superoxide) that are toxic to neurons and playa major role in the cellular immune response associated with the pathology of Alzheimer's disease(AD). OJaJiHwangEumJa(OJJHEJ) has been usually used for the treatment of senile disorders. For enhancing efficacy and convenience, the change of the drug delivery device of oriental herbal medicine is required. Objective: This experiment was designed to investigate the effect of the OJJHEJ hot water extract & ultra-fine powder on proinflammatory cytokine of microglia and memory deficit model. Method: The effects of the OJJHEJ hot water extract on production of IL-1$\beta$, IL-6, TNF-$\alpha$, in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. The effects of the OJJHEJ hot water extract & ultra-fine powder on the behavior of the memory deficit mice induced by scopolamine and AChE in serum of the memory deficit mice induced by scopolamine were investigated. Results: 1. The OJJHEJ hot water extract suppressed the production of IL-1$\beta$, IL-6, TNF-$\alpha$ in BV2 microglial cell line and the production of IL-6 was suppressed significantly. 2. The OJJHEJ hot water extract & ultra-fine powder decreased AChE significantly in the serum of the memory deficit mice induced by scopolamine. 3. The OJJHEJ hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. Conclusions: This experiment shows that the OJJHEJ hot water extract & ultra-fine powder might be effective for the prevention and treatment of memory impairment diseases. Investigation into the clinical use of the OJJHEJ hot water extract & ultra-fine powder for Alzheimer's disease is suggested for future research.

• 동의신경정신과학회지
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• 제20권1호
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• pp.59-73
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• 2009

Objectives : This experiment was designed to investigate the effect of the ODTCMT hot water extract & ultra-fine Powder on Alzheimer's Disease Model Induced by 13A. Methods : The effects of the ODTCMT hot water extract on expression of IL-l${\beta}$, IL-6, TNF-${\alpha}$ in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. The effects of the ODTCMT hot water extract & ultra-fine powder on the behavior of the memory deficit mice induced by scopolamine were investigated. Results : 1. The ODTCMT hot water extract significantly suppressed the production of TNF-${\alpha}$, IL-6 and IL-l${\beta}$ in BV2 microglial cell line treated with LPS. 2. The ODTCMT hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. Conclusions : These results suggest that the ODTCMT hot water extract & ultra-fine powder may be effective for the prevention and treatment of Alzheimer's disease. Investigation into the clinical use of the ODTCMT hot water extract & ultra-fine powder for Alzheimer's disease is suggested for future research.

• Archives of Pharmacal Research
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• 제28권2호
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• pp.216-219
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• 2005

Wogonin (5,7-dihydroxy-8-methoxyflavone) has been reported to exhibit a variety of biological properties including anti-inflammatory and neuroprotective functions. In this study, biological activities of diverse synthetic wogonin derivatives have been evaluated in two experimental cell culture models. Inhibitory activities of wogonin derivatives on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in BV2 microglial cells and on hydrogen peroxide ($H_{2}O_2$)-induced neuronal cell death in SH-SY5Y human neuroblastoma were examined. Wogonin derivatives such as WS2 and WS3 showed more potent suppressive activities on LPS-induced NO production and $H_{2}O_2$-induced cytotoxicity than wogonin itself. In addition, thiol substitution played a minor role in enhancing the activities of the derivatives. These findings may contribute to the development of novel anti-inflammatory and neuroprotective agents derived from wogonin.